- Peptide (C)RVGDAQGMFEPDGG, corresponding to amino acid residues 486-499 of rat NCX2 (Accession P48768). 3rd intracellular loop.
- Western blot analysis of rat brain membranes (lanes 1 and 5), mouse brain membranes (lanes 2 and 6), rat skeletal muscle lysate (lanes 3 and 7) and human SH-SY5Y neuroblastoma cell lysate (lanes 4 and 8):1-4. Anti-NCX2 (SLC8A2) Antibody (#ANX-012), (1:200).
5-8. Anti-NCX2 (SLC8A2) Antibody, preincubated with NCX2/SLC8A2 Blocking Peptide (#BLP-NX012).
- Expression of NCX2 in rat cerebellumImmunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NCX2 (SLC8A2) Antibody (#ANX-012), (1:100). A. NCX2 staining (red) is expressed in the molecular layer (M) and Purkinje cells (arrows). B. DAPI counterstain (blue) displays the layout of cerebellar layers. C. Merge of A and B demonstrates restriction of NCX2 expression to the molecular layer and Purkinje cells.
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Ca2+ has proven to be a universal signaling molecule in excitable and non-excitable cells. However, being that its intracellular concentration is 1000 times lower than that of the extracellular milieu, it is important for the cell to keep this ratio for proper function. NCX, a Na+/Ca2+ exchanger is responsible for most of the efflux of Ca2+ from the cell1-3. The NCX transporter is a member of the SLC8 family of solute carriers which in turn belong to the CaCA superfamily1,4,5.
NCX-1 is one of three Na+/Ca2+ exchangers (NCX-1, NCX-2, NCX-3) leading to one Ca2+ movement across the plasma membrane in exchange of three Na+ influx. However, the transporter can reverse the direction of the transport if the concentrations of Na+ and Ca2+ change6. The transporter has nine transmembrane domains and intracellular N- and C-terminals. Between tansmembrane domains 5 and 6, the presence of an extra-long intracellular loop, termed the f-loop, is responsible for regulating the activity of NCX-1 via several different mechanisms like ion binding, phosphorylation, etc. The f-loop also has sites which undergo alternative splicing7.
Of the three NCX expressed in mammalian cells, NCX-1 is the most widely expressed. Its expression is detected in the heart, brain, and kidney. NCX-1 undergoes alternative splicing in a tissue-dependent manner. The first splice region does not change the overall structure of the protein but rather enables the expression of the gene specific to the tissues which require the expression of the gene. The second splicing site leads to a number of proteins varying in length. NCX-2 expression is much more limited; it is expressed only in neurons. NCX-3 is expressed in skeletal muscle and in some regions of the brain and undergoes alternative splicing in a similar fashion to that of NCX-11,8.
Due to their central role in modulating Ca2+ levels in the cell, NCX exchangers are involved in various pathophysiological diseases/disorders such as hypoxia, aging, Alzheimer’s disease7, to name a few.
Species reactivity key:
Anti-NCX2 (SLC8A2) Antibody (#ANX-012) is a highly specific antibody directed against an epitope of the rat Na+/Ca2+ exchange protein 2. The antibody can be used in western blot ans immunohistochemistry applications. It has been designed to recognize NCX2 from human, rat, and mouse samples.
- Human glioblastoma LN-229, T98G and U373 cell lysates.
Song, M. et al. (2014) Br. J. Pharmacol. 171, 3432.